Using the Biofluorometer with Muscle Physiology Research systems

The use of fluorescence for sensing and imaging of the cellular signaling pathways has emerged as an indispensable tool in modern physiology, providing dynamic information of quantity and localization of the molecules of interest. Using appropriate indicator dyes, molecules alter their fluorescent characteristics in response to ion binding or membrane integration, so that the optical signal from the indicator can be measured to monitor the amplitude and the time course of various metal ions like Na+, K+, Mg2+ and Ca2+, as well as pH and membrane potential, in cellular compartments.

A specific target molecule like Ca2+ is responsible for many physiological functions, such as neurotransmitter release, fertilization and ion channel functions. Studying the cellular channel functions is directly related to the transient increase in the myoplasmic free calcium concentration (Δ[Ca2+]), as a key intermediate signaling event between excitation and contraction of muscle fibers. This makes it essential for the analysis of the force development in muscles.

The concomitant assessment of both parameters simultaneously is therefore critical in the evaluation and interpretation of the force development characteristics of single muscle fibers. Fluorescence techniques used in conjunction with muscle research systems (like WPI’s SI-HTB2 or SI-CTS200) to record muscle force has therefore become a standard technique in cardiac muscle and skeletal muscle physiology. WPI’s Biofluorometer (SI-BF-100) was specifically developed to monitor rapid changes, e.g. in Δ[Ca2+], using high-power LED modules, optical fiber combiner and highly sensitive photomultiplier modules to detect even very weak fluorescent signals at sample rates of 1,000 ratios/s.

The Biofluorometer opens a wide field in functional fluorescence research, by studying the fundamental and/or applied aspects of the underlying energetics and signaling aspects of muscle contraction. This is notably useful in:

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